GSM6729186: P2; Homo sapiens; RNA-Seq
Culture protocol: Cartilage tissues were transferred into a hypoxia chamber (4% O2) immediately after being removed from OA patients and minced finely. Collagenase II at concentrations of 2%, or  0.1% was applied for 0.5, 1, 2, 4, and 18 hours to digest the minced tissue.  Cell yield, viability, cell size, and transcriptome were determined using counter chamber, flow cytometry, and RNA sequencing (RNA-seq). During 4-hour digestions, collagenase II at 2% yielded maximal release, similar as 0.1% collagenase II-18 hour digestion. also of the large chondrocyte population RNA was isolated using RNeasy Mini Kit. RNA quality was determined on a Bioanalyzer using the RNA 6000 Pico Kit (Agilent). Illumina libraries were prepared using Smart-Seq v4 mRNA Ultra Low Input RNA Kit (Clontech) and Nextera XT DNA Sample Preparation Kit (Illumina), with 10 ng of purified cDNA, according to the manufacturer's instructions